Hello Hiren, (reply below)

On Tue, 2013-07-09 at 16:59 +0200, Hiren Ghosh wrote:
> Dear All, I used MIRA for genome assembly and i got contigfile "
> Ecoli-1_out.unpadded.fasta". Now i want to do alignment with this
> contig with another Ecoli genome " refseq.fasta". In command line i am
> typinglike this and getting error message. 
> Attempt 1: ./mauveAligner --output=ec_ec.mauve
> --output-alignment=ec_vs_ec.alignment Ecoli-1_out.unpadded.fasta
> refseq.fasta Sequence loaded successfully. Ecoli-1_out.unpadded.fasta
> 4982364 base pairs. Using weight 15 mers for initial seeds Creating
> sorted mer list Create time was: 2 seconds.
> 0%..1%..2%..3%..4%..5%..6%..7%..8%..9%..10%..
> 11%..12%..13%..14%..15%..16%..17%..18%..19%..20%..
> 21%..22%..23%..24%..25%..26%..27%..28%..29%..30%..
> 31%..32%..33%..34%..35%..36%..37%..38%..39%..40%..
> 41%..42%..43%..44%..45%..46%..47%..48%..49%..50%..
> 51%..52%..53%..54%..55%..56%..57%..58%..59%..60%..
> 61%..62%..63%..64%..65%..66%..67%..68%..69%..70%..
> 71%..72%..73%..74%..75%..76%..77%..78%..79%..80%..
> 81%..82%..83%..84%..85%..86%..87%..88%..89%..90%..
> 91%..92%..93%..94%..95%..96%..97%..98%..99%..Starting with 0 MUMs
> Eliminating overlaps yields 0 MUMs

In this case, mauveAligner was unable to find any exact matches of
length 15 or greater among your input sequences. This generally happens
when the genomes are too divergent to be aligned at the nucleotide
level. Unless you want mauveAligner for some historical reason, I
suggest you use progressiveMauve if possible since it can align more
divergent genomes than mauveAligner and is generally more accurate. 
However, if mauveAligner couldn't find any matches at all,
progressiveMauve will probably not do much better. Check that your
refseq input file does in fact contain sequence data -- there was only
one message in the output above giving the sequence size but that is
usually printed for both sequences.


> Attempt2: ./mauveAligner --output=jb50vsSpmito.mauve
> --output-alignment=jb50vsSpmito.alignment Ecoli-1_out.unpadded.fasta
> jb50contigs.fa.sslist refseq.fasta Sp.may2011.extras_mito.fasta.sslist
> Sequence loaded successfully. Ecoli-1_out.unpadded.fasta 4982364 base
> pairs. Sequence loaded successfully. refseq.fasta 2435000 base pairs.
> Using weight 15 mers for initial seeds Creating sorted mer list Create
> time was: 2 seconds. Creating sorted mer list ERROR! gap character
> encountered at genome sequence position 61 Input sequences must be
> unaligned and ungapped!

Unlike many errors generated by Mauve, this error message is pretty
self-explanatory. There is some kind of non-nucleotide character in your
input sequence. I think MIRA assembler output includes non-nucleotide
characters in some cases. Check the MIRA documentation and fix your
input files accordingly.

Best,
-Aaron

-- 
Aaron E. Darling, Ph.D.
Associate Professor, ithree institute
University of Technology Sydney
Australia

http://darlinglab.org
twitter: @koadman



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